Name: Late stage assay provider results from the probe development effort to identify inhibitors of PME-1: LC-MS/MS-based cell-based ABPP-SILAC assay. ..more
Depositor Specified Assays
Show more |
| AID | Name | Type | Probe | Comment |
|---|---|---|---|---|
| 2130 | Fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of Protein Phosphatase Methylesterase 1 (PME-1). | screening | Primary screen (PME-1 inhibitors) | |
| 2143 | Summary of probe development efforts to identify inhibitors of Protein Phosphatase Methylesterase 1 (PME-1). | summary | 3 | Summary (PME-1 inhibitors) |
| 2174 | Counterscreen for PME1 inhibitors: fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of lysophospholipase 1 (LYPLA1). | screening | Counterscreen (LYPLA1 inhibitors) | |
| 2171 | Fluorescence polarization-based biochemical high throughput confirmation assay for inhibitors of Protein Phosphatase Methylesterase 1 (PME-1). | screening | Confirmation assay (PME-1 inhibitors) | |
| 2177 | Counterscreen for PME1 inhibitors: fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of lysophospholipase 2 (LYPLA2). | screening | Counterscreen (LYPLA2 inhibitors) | |
| 2232 | Counterscreen for PME1 inhibitors: fluorescence polarization-based biochemical high throughput confirmation assay to identify inhibitors of lysophospholipase 2 (LYPLA2). | screening | Counterscreen confirmation (LYPLA2 inhibitors) | |
| 2233 | Counterscreen for PME1 inhibitors: fluorescence polarization-based biochemical high throughput confirmation assay for inhibitors of lysophospholipase 1 (LYPLA1). | screening | Counterscreen confirmation (LYPLA1 inhibitors) | |
| 2291 | Fluorescence polarization-based Maybridge primary biochemical high throughput screening assay to identify inhibitors of Protein Phosphatase Methylesterase 1 (PME-1). | screening | Primary screen (PME-1 inhibitors, Maybridge Library) | |
| 2363 | Late stage results from the probe development effort to identify inhibitors of the protein methylesterase PME-1: Inhibition of PME-1-mediated demethylation of PP2a | screening | MOA assay (Demethylation of PP2a) | |
| 2365 | Late stage results from the probe development effort to identify inhibitors of the protein methylesterase PME-1: Luminescence-based counterscreen assay to identify cytotoxic compounds | confirmatory | Counterscreen (Cytotoxicity HEC 293T) | |
| 2366 | Late stage results from the probe development effort to identify inhibitors of the protein methylesterase PME-1: Gel-based Activity-Based Protein Profiling (ABPP) IC50 | confirmatory | ABPP dose response screen (PME-1 inhibitors) | |
| 2368 | Late stage results from the probe development effort to identify inhibitors of the protein methylesterase PME-1: Gel-based Activity-Based Protein Profiling (ABPP) Gel Filtration Assay | screening | MOA assay (PME-1 inhibitors, gel filtration assay) | |
| 2369 | Late stage results from the probe development effort to identify inhibitors of the protein methylesterase PME-1: Gel-based Activity-Based Protein Profiling (ABPP) Inhibition | screening | ABPP screen (PME-1 inhibitors) | |
| 2371 | Late stage results from the probe development effort to identify inhibitors of the protein methylesterase PME-1: Gel-based Activity-Based Protein Profiling (ABPP) IC50: Purified enzyme | confirmatory | ABPP dose response screen (PME-1 inhibitors, purified enzyme) | |
| 463090 | Late stage assay provider results from the probe development effort to identify inhibitors of Protein Phosphatase Methylesterase 1 (PME-1): LC-MS/MS assay to assess binding of compounds to active site | other | 1 | MOA assay (PME-1 inhibitors, LC-MS assay) |
| 463091 | Late stage assay provider results from the probe development effort to identify inhibitors of Protein Phosphatase Methylesterase 1 (PME-1): luminescence-based biochemical dose response assay to determine cytotoxicity of inhibitor compounds | confirmatory | Counterscreen (Cytotoxicity HeLa) | |
| 463146 | Late stage assay provider results from the probe development effort to identify inhibitors of protein phosphatase methylesterase 1 (PME-1): Fluorescence-based biochemical gel-based ABPP | other | ABPP profiling (PME1 inhibitors in singlicate) | |
| 463149 | Late stage assay provider results from the probe development effort to identify inhibitors of PME-1: Fluorescence-based biochemical gel-based ABPP inhibition and selectivity | other | ABPP profiling (PME1 inhibition and selectivity in singlicate) | |
| 463131 | Late stage assay provider results from the probe development effort to identify inhibitors of protein phosphatase methylesterase 1 (PME-1): fluorescence-based cell-based inhibition | screening | ABPP profiling (PME1 inhibitors in singlicate) | |
| 463130 | Late stage assay provider results from the probe development effort to identify inhibitors of protein phosphatase methylesterase 1 (PME-1): Gel-based Activity-Based Protein Profiling (ABPP) IC50 Set 1 | confirmatory | ABPP profiling dose response (PME1 inhibitors in triplicate) | |
| 463124 | Late stage assay provider results from the probe development effort to identify inhibitors of protein phosphatase methylesterase 1 (PME-1): Gel-based Activity-Based Protein Profiling (ABPP) IC50 Set 2 | confirmatory | ABPP profiling dose response (PME1 inhibitors in triplicate) | |
| 463132 | Late stage assay provider results from the probe development effort to identify inhibitors of protein phosphatase methylesterase 1 (PME-1): Inhibition of PME-1-mediated demethylation of PP2A | screening | MOA assay (PP2a demethylation) | |
| 588835 | Late stage assay provider results from the probe development effort to identify inhibitors of PME-1: fluorescence-based dose response cell-based gel-based competitive Activity-Based Protein Profiling (ABPP) ABHD10 selectivity assay | other | ||
| 602468 | Late stage assay provider results from the probe development effort to identify inhibitors of PME-1: fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition and selectivity among cysteine-reactive proteins | other | ||
| 602485 | Late stage assay provider results from the probe development effort to identify inhibitors of PME-1: fluorescence-based cell-based gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of ABHD10 in vivo | other |
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Benjamin Cravatt, TSRI
Network: Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R01 CA132630-01
Grant Proposal PI: Benjamin Cravatt, TSRI
External Assay ID: PME1_INH_LCMS_ABPP-SILAC
Name: Late stage assay provider results from the probe development effort to identify inhibitors of PME-1: LC-MS/MS-based cell-based ABPP-SILAC assay.
Description:
Reversible protein phosphorylation networks play essential roles in most cellular processes. While over 500 kinases catalyze protein phosphorylation, only two enzymes, PP1 and PP2A, are responsible for > 90% of all serine/ threonine phosphatase activity (1). Phosphatases, unlike kinases, achieve substrate specificity through complex subunit assembly and post-translational modifications rather than number. PP2A, for example, typically exists as heterotrimer with diverse subunits that may combinatorially make as many as 70 different holoenzyme assemblies (2). Mutations in several of these PP2A subunits have been identified in human cancers, suggesting that PP2A may act as a tumor suppressor (3). Adding further complexity, several residues of the catalytic subunit of PP2A can be reversibly phosphorylated, and the C-terminal leucine residue can be reversibly methylated (4,5). PME-1 is specifically responsible for demethylation of the carboxyl terminus (6).
Methylesterification is thought to control the binding of different subunits to PP2A, but little is known about physiological significance of this post-translational modification in vivo (7). Recently, PME-1 has been identified as a protector of sustained ERK pathway activity in malignant gliomas (8). In order to further elucidate the role of PP2A methylation in vivo, our lab has generated mice that lack PME-1 (PME-1 (-/-) mice) by targeted gene disruption (9). Unfortunately, PME-1 deletion resulted in perinatal lethality, underscoring the importance of PME-1 but hindering our biological studies. Biochemical elucidation of PME-1 would thus greatly benefit from the development of potent and selective chemical inhibitors.
References:
1. Oliver, C. J., Shenolikar, S. (1998). Physiologic importance of protein phosphatase inhibitors. Front. Biosci. 3, D961-972.
2. Janssens, V., Goris, J. (2001). Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem. J. 353, 417-439.
3. Janssens, V., Goris, J., Van Hoof, C. (2005). PP2A: the expected tumor suppressor. Curr. Opin. Genet. Dev. 15, 34-41.
4. Chen, J., Martin, B. L., Brautigan, D. L. (1992). Regulation of protein serine-threonine phosphatase type-2A by tyrosine phosphorylation. Science 257, 1261-1264.
5. Favre, B., Zolnierowicz, S., Turowski, P., Hemmings, B. A. (1994). The catalytic subunit of protein phosphatase 2A is carboxyl-methylated in vivo. J. Biol. Chem. 269, 16311-16317.
6. Lee, J., Chen, Y., Tolstykh, T., Stock, J. (1996). A specific protein carboxyl methylesterase that demethylates phosphoprotein phosphatase 2A in bovine brain. Proc. Natl. Acad. Sci. U. S. A. 93, 6043-6047. .
7. Wu, J., Tolstykh, T., Lee, J., Boyd, K., Stock, J. B., Broach, J. R. (2000). Carboxyl methylation of the phosphoprotein phosphatase 2A catalytic subunit promotes its functional association with regulatory subunits in vivo. Embo J. 19, 5672-5681.
8. Puustinen, P., Junttila, M. R., Vanhatupa, S., Sablina, A. A., Hector, M. E., Teittinen, K., Raheem, O., Ketola, K., Lin, S., Kast, J., Haapasalo, H., Hahn, W. C., Westermarck, J. (2009). PME-1 protects extracellular signal-regulated kinase pathway activity from protein phosphatase 2A-mediated inactivation in human malignant glioma. Cancer Res. 69, 2870-2877.
9. Ortega-Gutierrez, S., Leung, D., Ficarro, S., Peters, E. C., Cravatt, B. F. (2008). Targeted disruption of the PME-1 gene causes loss of demethylated PP2A and perinatal lethality in mice. PLoS ONE 3, e2486.
Keywords:
late stage, late stage AID, assay provider, powders, PME-1, protein phosphatase methylesterase 1, PPME-1, protein phosphatase 2a, PP2A, methylation, demethylation, counterscreen, liquid chromatography, LC, tandem mass spectrometry, MS/MS, activity-based protein profiling, ABPP, stable isotope labeling with amino acids in cell culture, SILAC, ABPP-SILAC, fluorophosphonate biotin, FP-Rh, inhibitor, selectivity, anti-targets, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Benjamin Cravatt, TSRI
Network: Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R01 CA132630-01
Grant Proposal PI: Benjamin Cravatt, TSRI
External Assay ID: PME1_INH_LCMS_ABPP-SILAC
Name: Late stage assay provider results from the probe development effort to identify inhibitors of PME-1: LC-MS/MS-based cell-based ABPP-SILAC assay.
Description:
Reversible protein phosphorylation networks play essential roles in most cellular processes. While over 500 kinases catalyze protein phosphorylation, only two enzymes, PP1 and PP2A, are responsible for > 90% of all serine/ threonine phosphatase activity (1). Phosphatases, unlike kinases, achieve substrate specificity through complex subunit assembly and post-translational modifications rather than number. PP2A, for example, typically exists as heterotrimer with diverse subunits that may combinatorially make as many as 70 different holoenzyme assemblies (2). Mutations in several of these PP2A subunits have been identified in human cancers, suggesting that PP2A may act as a tumor suppressor (3). Adding further complexity, several residues of the catalytic subunit of PP2A can be reversibly phosphorylated, and the C-terminal leucine residue can be reversibly methylated (4,5). PME-1 is specifically responsible for demethylation of the carboxyl terminus (6).
Methylesterification is thought to control the binding of different subunits to PP2A, but little is known about physiological significance of this post-translational modification in vivo (7). Recently, PME-1 has been identified as a protector of sustained ERK pathway activity in malignant gliomas (8). In order to further elucidate the role of PP2A methylation in vivo, our lab has generated mice that lack PME-1 (PME-1 (-/-) mice) by targeted gene disruption (9). Unfortunately, PME-1 deletion resulted in perinatal lethality, underscoring the importance of PME-1 but hindering our biological studies. Biochemical elucidation of PME-1 would thus greatly benefit from the development of potent and selective chemical inhibitors.
References:
1. Oliver, C. J., Shenolikar, S. (1998). Physiologic importance of protein phosphatase inhibitors. Front. Biosci. 3, D961-972.
2. Janssens, V., Goris, J. (2001). Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem. J. 353, 417-439.
3. Janssens, V., Goris, J., Van Hoof, C. (2005). PP2A: the expected tumor suppressor. Curr. Opin. Genet. Dev. 15, 34-41.
4. Chen, J., Martin, B. L., Brautigan, D. L. (1992). Regulation of protein serine-threonine phosphatase type-2A by tyrosine phosphorylation. Science 257, 1261-1264.
5. Favre, B., Zolnierowicz, S., Turowski, P., Hemmings, B. A. (1994). The catalytic subunit of protein phosphatase 2A is carboxyl-methylated in vivo. J. Biol. Chem. 269, 16311-16317.
6. Lee, J., Chen, Y., Tolstykh, T., Stock, J. (1996). A specific protein carboxyl methylesterase that demethylates phosphoprotein phosphatase 2A in bovine brain. Proc. Natl. Acad. Sci. U. S. A. 93, 6043-6047. .
7. Wu, J., Tolstykh, T., Lee, J., Boyd, K., Stock, J. B., Broach, J. R. (2000). Carboxyl methylation of the phosphoprotein phosphatase 2A catalytic subunit promotes its functional association with regulatory subunits in vivo. Embo J. 19, 5672-5681.
8. Puustinen, P., Junttila, M. R., Vanhatupa, S., Sablina, A. A., Hector, M. E., Teittinen, K., Raheem, O., Ketola, K., Lin, S., Kast, J., Haapasalo, H., Hahn, W. C., Westermarck, J. (2009). PME-1 protects extracellular signal-regulated kinase pathway activity from protein phosphatase 2A-mediated inactivation in human malignant glioma. Cancer Res. 69, 2870-2877.
9. Ortega-Gutierrez, S., Leung, D., Ficarro, S., Peters, E. C., Cravatt, B. F. (2008). Targeted disruption of the PME-1 gene causes loss of demethylated PP2A and perinatal lethality in mice. PLoS ONE 3, e2486.
Keywords:
late stage, late stage AID, assay provider, powders, PME-1, protein phosphatase methylesterase 1, PPME-1, protein phosphatase 2a, PP2A, methylation, demethylation, counterscreen, liquid chromatography, LC, tandem mass spectrometry, MS/MS, activity-based protein profiling, ABPP, stable isotope labeling with amino acids in cell culture, SILAC, ABPP-SILAC, fluorophosphonate biotin, FP-Rh, inhibitor, selectivity, anti-targets, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN
Panel Information
Targets
§ Panel component ID.
| Data Table(Active) Data Table(All) | Show more |
| PID§ | Name | Substance | Panel Targets | Description | Additional Information | ||
|---|---|---|---|---|---|---|---|
| Active | Inactive | ||||||
| 1 | Pme1 | 1 | protein phosphatase methylesterase 1 [Mus musculus] [gi:30794138] | Taxonomy id: 10090 Gene id: 72590 | |||
| 2 | Abhd10 | 1 | abhydrolase domain-containing protein 10, mitochondrial precursor [Mus musculus] [gi:269784760] | Taxonomy id: 10090 Gene id: 213012 | |||
| 3 | Abhd11 | 1 | abhydrolase domain-containing protein 11 isoform 1 [Mus musculus] [gi:21644577] | Taxonomy id: 10090 Gene id: 68758 | |||
| 4 | Abhd12 | 1 | monoacylglycerol lipase ABHD12 [Mus musculus] [gi:159110817] | Taxonomy id: 10090 Gene id: 76192 | |||
| 5 | Abhd6 | 1 | monoacylglycerol lipase ABHD6 [Mus musculus] [gi:31560264] | Taxonomy id: 10090 Gene id: 66082 | |||
| 6 | Ache | 1 | acetylcholinesterase precursor [Mus musculus] [gi:13928664] | Taxonomy id: 10090 Gene id: 11423 | |||
| 7 | Acot1 | 1 | acyl-coenzyme A thioesterase 1 [Mus musculus] [gi:6753550] | Taxonomy id: 10090 Gene id: 26897 | |||
| 8 | Acot2 | 1 | acyl-coenzyme A thioesterase 2, mitochondrial precursor [Mus musculus] [gi:238624114] | Taxonomy id: 10090 Gene id: 171210 | |||
| 9 | Acot7 | 1 | cytosolic acyl coenzyme A thioester hydrolase isoform 1 [Mus musculus] [gi:225690616] | Taxonomy id: 10090 Gene id: 70025 | |||
| 10 | Acot9 | 1 | acyl-coenzyme A thioesterase 9, mitochondrial [Mus musculus] [gi:31980998] | Taxonomy id: 10090 Gene id: 56360 | |||
| 11 | Apeh | 1 | Acylpeptide hydrolase [Mus musculus] [gi:19343726] | Taxonomy id: 10090 Gene id: 235606 | |||
| 12 | Bat5 | 1 | abhydrolase domain-containing protein 16A [Mus musculus] [gi:30519896] | Taxonomy id: 10090 Gene id: 193742 | |||
| 13 | Ctsa | 1 | lysosomal protective protein isoform b [Mus musculus] [gi:84042523] | Taxonomy id: 10090 Gene id: 19025 | |||
| 14 | Ddhd2 | 1 | phospholipase DDHD2 [Mus musculus] [gi:254692989] | Taxonomy id: 10090 Gene id: 72108 | |||
| 15 | Dpp7 | 1 | dipeptidyl peptidase 2 precursor [Mus musculus] [gi:31981425] | Taxonomy id: 10090 Gene id: 83768 | |||
| 16 | Dpp8 | 1 | dipeptidyl peptidase 8 [Mus musculus] [gi:31542571] | Taxonomy id: 10090 Gene id: 74388 | |||
| 17 | Dpp9 | 1 | dipeptidyl peptidase 9 [Mus musculus] [gi:255003757] | Taxonomy id: 10090 Gene id: 224897 | |||
| 18 | Faah | 1 | fatty acid amide hydrolase [Mus musculus] [gi:123253900] | Taxonomy id: 10090 Gene id: 14073 | |||
| 19 | Fam108b1 | 1 | abhydrolase domain-containing protein FAM108B1 precursor [Mus musculus] [gi:38142456] | Taxonomy id: 10090 Gene id: 226016 | |||
| 20 | Fasn | 1 | fatty acid synthase [Mus musculus] [gi:93102409] | Taxonomy id: 10090 Gene id: 14104 | |||
| 21 | Htra2 | 1 | serine protease HTRA2, mitochondrial [Mus musculus] [gi:254281222] | Taxonomy id: 10090 Gene id: 64704 | |||
| 22 | Iah1 | 1 | isoamyl acetate-hydrolyzing esterase 1 homolog [Mus musculus] [gi:27754071] | Taxonomy id: 10090 Gene id: 67732 | |||
| 23 | Lypla1 | 1 | acyl-protein thioesterase 1 [Mus musculus] [gi:6678760] | Taxonomy id: 10090 Gene id: 18777 | |||
| 24 | Lypla2 | 1 | acyl-protein thioesterase 2 [Mus musculus] [gi:7242156] | Taxonomy id: 10090 Gene id: 26394 | |||
| 25 | Lyplal1 | 1 | lysophospholipase-like protein 1 [Mus musculus] [gi:227496223] | Taxonomy id: 10090 Gene id: 226791 | |||
| 26 | Nceh1 | 1 | neutral cholesterol ester hydrolase 1 [Mus musculus] [gi:30520239] | Taxonomy id: 10090 Gene id: 320024 | |||
| 27 | Pafah1b2 | 1 | platelet-activating factor acetylhydrolase IB subunit beta [Mus musculus] [gi:40254624] | Taxonomy id: 10090 Gene id: 18475 | |||
| 28 | Pafah1b3 | 1 | platelet-activating factor acetylhydrolase IB subunit gamma [Mus musculus] [gi:6679201] | Taxonomy id: 10090 Gene id: 18476 | |||
| 29 | Pafah2 | 1 | platelet-activating factor acetylhydrolase 2, cytoplasmic [Mus musculus] [gi:225579137] | Taxonomy id: 10090 Gene id: 100163 | |||
| 30 | Parl | 1 | presenilins-associated rhomboid-like protein, mitochondrial precursor [Mus musculus] [gi:54261813] | Taxonomy id: 10090 Gene id: 381038 | |||
| 31 | Pgap1 | 1 | GPI inositol-deacylase [Mus musculus] [gi:254028203] | Taxonomy id: 10090 Gene id: 241062 | |||
| 32 | Pla2g15 | 1 | group XV phospholipase A2 precursor [Mus musculus] [gi:19527008] | Taxonomy id: 10090 Gene id: 192654 | |||
| 33 | Pla2g6 | 1 | 85 kDa calcium-independent phospholipase A2 isoform 2 [Mus musculus] [gi:312222739] | Taxonomy id: 10090 Gene id: 53357 | |||
| 34 | Pnpla8 | 1 | calcium-independent phospholipase A2-gamma [Mus musculus] [gi:118130807] | Taxonomy id: 10090 Gene id: 67452 | |||
| 35 | Ppt2 | 1 | lysosomal thioesterase PPT2 precursor [Mus musculus] [gi:9506985] | Taxonomy id: 10090 Gene id: 54397 | |||
| 36 | Prcp | 1 | lysosomal Pro-X carboxypeptidase precursor [Mus musculus] [gi:33469015] | Taxonomy id: 10090 Gene id: 72461 | |||
| 37 | Prep | 1 | prolyl endopeptidase [Mus musculus] [gi:6755152] | Taxonomy id: 10090 Gene id: 19072 | |||
| 38 | Prepl | 1 | prolyl endopeptidase-like isoform a [Mus musculus] [gi:254939518] | Taxonomy id: 10090 Gene id: 213760 | |||
| 39 | Rbbp9 | 1 | putative hydrolase RBBP9 [Mus musculus] [gi:86439977] | Taxonomy id: 10090 Gene id: 26450 | |||
| 40 | Serhl | 1 | serine hydrolase-like protein [Mus musculus] [gi:13443008] | Taxonomy id: 10090 Gene id: 68607 | |||
§ Panel component ID.
Protocol
Assay Overview:
The purpose of this assay is to determine the selectivity profile of powder samples of test compounds using activity-based protein profiling (ABPP) in combination with stable isotope labeling with amino acids in cell culture (SILAC). In this assay, cultured Neuro-2A cells are metabolically labeled with light or heavy amino acids. Light and heavy cells are treated with test compound and DMSO, respectively, in situ. Cells are lysed, proteomes are treated with the serine-hydrolase-specific activity-based fluorophosphonate-biotin (FP-biotin) affinity probe, and combined in a 1:1 (w/w) ratio. Biotinylated proteins are enriched, trypsinized, and analyzed by multi-dimensional liquid chromatography tandem mass spectrometery LC/LC-MS/MS (MudPIT). Inhibition of target and anti-target activity is quantified by comparing intensities of light and heavy peptide peaks. As designed, compounds that act as inhibitors will block FP-biotin labeling, reducing enrichment in the inhibitor-treated (light) sample relative to the DMSO-treated (heavy) sample, resulting in a smaller light/heavy ratio for each protein. Proteins not targeted by inhibitors would be expected to have a ratio of 1.
Protocol Summary:
Stable isotope labeling with amino acids in cell culture (SILAC). Neuro-2A murine neuroblastoma cells were initially grown for 10 passages in either light or heavy SILAC DMEM medium supplemented with 10% dialyzed FCS and 2 mM L-glutamine. Light media was supplemented with 100 ug/mL L-arginine and 100 ug/mL L-lysine. Heavy media was supplemented with 100 ug/mL [13C615N4]-L-Arginine and 100 ug/mL [13C615N2]-L-Lysine. Light cells (in 10 mL media) were treated with 100 nM test compound (75 uL of a 200x stock in DMSO) and heavy cells were treated with DMSO (75 uL) for 2 hours at 37 C. Cells were washed 2 times with DPBS, harvested, and homogenized by sonication in DPBS (1mL). The soluble and membrane fractions were isolated by centrifugation (100K x g, 45 minutes) and the protein concentration was adjusted to 2 mg/mL with DPBS in each fraction.
Sample preparation for ABPP-SILAC. The light and heavy proteomes were labeled with 10 uM of FP-biotin (500 uL total reaction volume) for 2 hours at 25 C. After incubation, light and heavy proteomes were mixed in 1:1 ratio, and the membrane proteomes were additionally solubilized with 1% Triton-X100. The proteomes were desalted over PD-10 desalting columns (GE Healthcare) and FP-labeled proteins were enriched with streptavidin beads. The beads were washed with 1% SDS in DPBS (1x), 6M urea (1x), and DPBS (2x), then resuspended in 6 M urea, reduced with 5 mM TCEP for 20 minutes, and alkylated with 10 mM iodoacetamide for 30 minutes at 25 C in the dark. On-bead digestions were performed for 12 hours at 37 C with sequence-grade modified trypsin (Promega; 2 ug) in 2M urea in the presence of 2 mM CaCl2. Peptide samples were acidified to a final concentration of 5% (v/v) formic acid, pressure-loaded on to a biphasic (strong cation exchange/reversed phase) capillary column and analyzed as described below.
LC-MS/MS analysis. Digested and acidified peptide mixtures were analyzed by two-dimensional liquid chromatography (2D-LC) separation in combination with tandem mass spectrometry using an Agilent 1200-series quaternary pump and Thermo Scientific LTQ-Orbitrap Velos ion trap mass spectrometer. Peptides were eluted in a 5-step MudPIT experiment using 0%, 25%, 50%, 80%, and 100% salt bumps of 500 mM aqueous ammonium acetate and data were collected in data-dependent acquisition mode with dynamic exclusion turned on (20 s, repeat of 1). Specifically, one full MS (MS1) scan (400-1800 m/z) was followed by 30 MS2 scans of the most abundant ions. The MS2 spectra data were extracted from the raw file using RAW Xtractor (version 1.9.9.2; publicly available at http://fields.scripps.edu/downloads.php). MS2 spectra data were searched using the ProLuCID algorithm (publicly available at http://fields.scripps.edu/downloads.php) against the latest version of the mouse IPI database concatenated with the reversed database for assessment of false-discovery rates. ProLucid searches allowed for static modification of cysteine residues (+57.02146 due to alkylation), methionine oxidation (+15.9949), mass shifts of labeled amino acids (+10.0083 R, 8.0142 K) and no enzyme specificity. The resulting MS2 spectra matches were assembled into protein identifications and filtered using DTASelect (version 2.0) using the --modstat, --mass, and --trypstat options (applies different statistical models for the analysis of high resolution masses, peptide digestion state, and methionine oxidation state respectively). Ratios of Light/Heavy peaks were calculated using in-house software and normalized at the peptide level to the average ratio of all non-serine hydrolase peptides. Reported ratios represent the mean of all unique, quantified peptides per protein and do not include peptides that were greater than 3 standard deviations from the median peptide value. Proteins with less than three peptides per protein ID were not included in the analysis.
Ratio = Average( AUC_light / AUC_heavy ) calculated for all unique peptides
Where:
AUC_light is the area-under-the-curve for the light peptide pair from cells treated with test compound.
AUC_heavy is the area-under-the-curve for the heavy peptide pair from cells treated with DMSO.
PubChem Activity Outcome and Score:
The following applies to each panel:
A compound with a light/heavy ratio of less than or equal to 0.5 for a particular target/anti-target was considered "active". A compound with a light/heavy ratio of greater than 0.5 for a specified target/anti-target was considered "inactive".
Overall Outcome and Score:
A compound was considered active if it was active for PME1 and inactive for all anti-target serine hydrolases tested.
The PubChem Activity Score is assigned a value of 100 for active compounds, and 0 for inactive compounds.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
Neuro-2A murine cells (provided by Assay Provider)
SILAC DMEM media (Thermo 89985)
dialyzed FCS (Gemini 100-108)
1x Glutamine (CellGro 25-005-CI)
L-Arginine (SigmaAldrich A6969)
L-Lysine (SigmaAldrich L9037)
[13C615N4]-L-Arginine (SigmaAldrich 608033)
[13C615N2]-L-Lysine (SigmaAldrich 608041)
DPBS (Cellgro 20-031-CV)
FP-biotin (provided by Assay Provider)
PD-10 desalting columns (GE Healthcare 17-0851-01)
SDS (SigmaAldrich L6026)
Urea (Fisher U15-3)
TCEP (SigmaAldrich 75259)
Iodoacetamide (SigmaAldrich I1149)
Trypsin (Promega V5111)
CaCl2 (SigmaAldrich C1016)
Streptavidin beads (Pierce 20349)
Fused-silica (Agilent 160-2635-10)
Strong cation exchange (Phenomenex CH0-2257)
Aqua C18 (Phenomenex 04A-4299)
Acetonitrile (Fisher A955-4)
Millipore-filtered Water
Formic acid (Fluka 06440)
Triton-X100 (Fisher AC21568-0010)
The purpose of this assay is to determine the selectivity profile of powder samples of test compounds using activity-based protein profiling (ABPP) in combination with stable isotope labeling with amino acids in cell culture (SILAC). In this assay, cultured Neuro-2A cells are metabolically labeled with light or heavy amino acids. Light and heavy cells are treated with test compound and DMSO, respectively, in situ. Cells are lysed, proteomes are treated with the serine-hydrolase-specific activity-based fluorophosphonate-biotin (FP-biotin) affinity probe, and combined in a 1:1 (w/w) ratio. Biotinylated proteins are enriched, trypsinized, and analyzed by multi-dimensional liquid chromatography tandem mass spectrometery LC/LC-MS/MS (MudPIT). Inhibition of target and anti-target activity is quantified by comparing intensities of light and heavy peptide peaks. As designed, compounds that act as inhibitors will block FP-biotin labeling, reducing enrichment in the inhibitor-treated (light) sample relative to the DMSO-treated (heavy) sample, resulting in a smaller light/heavy ratio for each protein. Proteins not targeted by inhibitors would be expected to have a ratio of 1.
Protocol Summary:
Stable isotope labeling with amino acids in cell culture (SILAC). Neuro-2A murine neuroblastoma cells were initially grown for 10 passages in either light or heavy SILAC DMEM medium supplemented with 10% dialyzed FCS and 2 mM L-glutamine. Light media was supplemented with 100 ug/mL L-arginine and 100 ug/mL L-lysine. Heavy media was supplemented with 100 ug/mL [13C615N4]-L-Arginine and 100 ug/mL [13C615N2]-L-Lysine. Light cells (in 10 mL media) were treated with 100 nM test compound (75 uL of a 200x stock in DMSO) and heavy cells were treated with DMSO (75 uL) for 2 hours at 37 C. Cells were washed 2 times with DPBS, harvested, and homogenized by sonication in DPBS (1mL). The soluble and membrane fractions were isolated by centrifugation (100K x g, 45 minutes) and the protein concentration was adjusted to 2 mg/mL with DPBS in each fraction.
Sample preparation for ABPP-SILAC. The light and heavy proteomes were labeled with 10 uM of FP-biotin (500 uL total reaction volume) for 2 hours at 25 C. After incubation, light and heavy proteomes were mixed in 1:1 ratio, and the membrane proteomes were additionally solubilized with 1% Triton-X100. The proteomes were desalted over PD-10 desalting columns (GE Healthcare) and FP-labeled proteins were enriched with streptavidin beads. The beads were washed with 1% SDS in DPBS (1x), 6M urea (1x), and DPBS (2x), then resuspended in 6 M urea, reduced with 5 mM TCEP for 20 minutes, and alkylated with 10 mM iodoacetamide for 30 minutes at 25 C in the dark. On-bead digestions were performed for 12 hours at 37 C with sequence-grade modified trypsin (Promega; 2 ug) in 2M urea in the presence of 2 mM CaCl2. Peptide samples were acidified to a final concentration of 5% (v/v) formic acid, pressure-loaded on to a biphasic (strong cation exchange/reversed phase) capillary column and analyzed as described below.
LC-MS/MS analysis. Digested and acidified peptide mixtures were analyzed by two-dimensional liquid chromatography (2D-LC) separation in combination with tandem mass spectrometry using an Agilent 1200-series quaternary pump and Thermo Scientific LTQ-Orbitrap Velos ion trap mass spectrometer. Peptides were eluted in a 5-step MudPIT experiment using 0%, 25%, 50%, 80%, and 100% salt bumps of 500 mM aqueous ammonium acetate and data were collected in data-dependent acquisition mode with dynamic exclusion turned on (20 s, repeat of 1). Specifically, one full MS (MS1) scan (400-1800 m/z) was followed by 30 MS2 scans of the most abundant ions. The MS2 spectra data were extracted from the raw file using RAW Xtractor (version 1.9.9.2; publicly available at http://fields.scripps.edu/downloads.php). MS2 spectra data were searched using the ProLuCID algorithm (publicly available at http://fields.scripps.edu/downloads.php) against the latest version of the mouse IPI database concatenated with the reversed database for assessment of false-discovery rates. ProLucid searches allowed for static modification of cysteine residues (+57.02146 due to alkylation), methionine oxidation (+15.9949), mass shifts of labeled amino acids (+10.0083 R, 8.0142 K) and no enzyme specificity. The resulting MS2 spectra matches were assembled into protein identifications and filtered using DTASelect (version 2.0) using the --modstat, --mass, and --trypstat options (applies different statistical models for the analysis of high resolution masses, peptide digestion state, and methionine oxidation state respectively). Ratios of Light/Heavy peaks were calculated using in-house software and normalized at the peptide level to the average ratio of all non-serine hydrolase peptides. Reported ratios represent the mean of all unique, quantified peptides per protein and do not include peptides that were greater than 3 standard deviations from the median peptide value. Proteins with less than three peptides per protein ID were not included in the analysis.
Ratio = Average( AUC_light / AUC_heavy ) calculated for all unique peptides
Where:
AUC_light is the area-under-the-curve for the light peptide pair from cells treated with test compound.
AUC_heavy is the area-under-the-curve for the heavy peptide pair from cells treated with DMSO.
PubChem Activity Outcome and Score:
The following applies to each panel:
A compound with a light/heavy ratio of less than or equal to 0.5 for a particular target/anti-target was considered "active". A compound with a light/heavy ratio of greater than 0.5 for a specified target/anti-target was considered "inactive".
Overall Outcome and Score:
A compound was considered active if it was active for PME1 and inactive for all anti-target serine hydrolases tested.
The PubChem Activity Score is assigned a value of 100 for active compounds, and 0 for inactive compounds.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
Neuro-2A murine cells (provided by Assay Provider)
SILAC DMEM media (Thermo 89985)
dialyzed FCS (Gemini 100-108)
1x Glutamine (CellGro 25-005-CI)
L-Arginine (SigmaAldrich A6969)
L-Lysine (SigmaAldrich L9037)
[13C615N4]-L-Arginine (SigmaAldrich 608033)
[13C615N2]-L-Lysine (SigmaAldrich 608041)
DPBS (Cellgro 20-031-CV)
FP-biotin (provided by Assay Provider)
PD-10 desalting columns (GE Healthcare 17-0851-01)
SDS (SigmaAldrich L6026)
Urea (Fisher U15-3)
TCEP (SigmaAldrich 75259)
Iodoacetamide (SigmaAldrich I1149)
Trypsin (Promega V5111)
CaCl2 (SigmaAldrich C1016)
Streptavidin beads (Pierce 20349)
Fused-silica (Agilent 160-2635-10)
Strong cation exchange (Phenomenex CH0-2257)
Aqua C18 (Phenomenex 04A-4299)
Acetonitrile (Fisher A955-4)
Millipore-filtered Water
Formic acid (Fluka 06440)
Triton-X100 (Fisher AC21568-0010)
Comment
This assay was performed by the assay provider with powder samples of synthetic compounds.
Categorized Comment
BAO: version: 1.4b1090
BAO: bioassay specification: assay stage: secondary: selectivity
BAO: bioassay specification: assay biosafety level: bsl1
BAO: assay format: biochemical format: protein format: protein complex format
BAO: bioassay specification: assay measurement type: endpoint assay
BAO: bioassay specification: assay readout content: assay readout method: regular screening
BAO: bioassay specification: assay readout content: content readout type: single readout
BAO: meta target: molecular target: protein target: enzyme: generic hydrolase
BAO: meta target detail: binding reporter specification: interaction: protein-small molecule
BAO: bioassay specification: assay stage: secondary: selectivity
BAO: bioassay specification: assay biosafety level: bsl1
BAO: assay format: biochemical format: protein format: protein complex format
BAO: bioassay specification: assay measurement type: endpoint assay
BAO: bioassay specification: assay readout content: assay readout method: regular screening
BAO: bioassay specification: assay readout content: content readout type: single readout
BAO: meta target: molecular target: protein target: enzyme: generic hydrolase
BAO: meta target detail: binding reporter specification: interaction: protein-small molecule
Result Definitions
| Show more |
| TID | Name | Description | PID§ | Panel Targets | Histogram | Type | Unit | |
|---|---|---|---|---|---|---|---|---|
| Outcome | The BioAssay activity outcome | Outcome | ||||||
| Score | The BioAssay activity ranking score |  | Integer | |||||
| 1 | Outcome [Pme1] | One of Active, Inactive, or Not Tested | 1 | protein phosphatase methylesterase 1 [Mus musculus] | | Outcome | ||
| 2 | Mean Ratio [Pme1] | Calculated inhibitor-treated/DMSO ratio for Pme1 as mean for all unique peptides quantified | 1 | | Float | ratio | ||
| 3 | Standard Error [Pme1] | Calculated standard error for mean | 1 | | Float | |||
| 4 | Outcome [Abhd10] | One of Active, Inactive, or Not Tested | 2 | abhydrolase domain-containing protein 10, mitochondrial precursor [Mus musculus] | | Outcome | ||
| 5 | Mean Ratio [Abhd10] | Calculated inhibitor-treated/DMSO ratio for Abhd10 as mean for all unique peptides quantified | 2 | | Float | ratio | ||
| 6 | Standard Error [Abhd10] | Calculated standard error for mean | 2 | | Float | |||
| 7 | Outcome [Abhd11] | One of Active, Inactive, or Not Tested | 3 | abhydrolase domain-containing protein 11 isoform 1 [Mus musculus] | | Outcome | ||
| 8 | Mean Ratio [Abhd11] | Calculated inhibitor-treated/DMSO ratio for Abhd11 as mean for all unique peptides quantified | 3 | | Float | ratio | ||
| 9 | Standard Error [Abhd11] | Calculated standard error for mean | 3 | | Float | |||
| 10 | Outcome [Abhd12] | One of Active, Inactive, or Not Tested | 4 | monoacylglycerol lipase ABHD12 [Mus musculus] | | Outcome | ||
| 11 | Mean Ratio [Abhd12] | Calculated inhibitor-treated/DMSO ratio for Abhd12 as mean for all unique peptides quantified | 4 | | Float | ratio | ||
| 12 | Standard Error [Abhd12] | Calculated standard error for mean | 4 | | Float | |||
| 13 | Outcome [Abhd6] | One of Active, Inactive, or Not Tested | 5 | monoacylglycerol lipase ABHD6 [Mus musculus] | | Outcome | ||
| 14 | Mean Ratio [Abhd6] | Calculated inhibitor-treated/DMSO ratio for Abhd6 as mean for all unique peptides quantified | 5 | | Float | ratio | ||
| 15 | Standard Error [Abhd6] | Calculated standard error for mean | 5 | | Float | |||
| 16 | Outcome [Ache] | One of Active, Inactive, or Not Tested | 6 | acetylcholinesterase precursor [Mus musculus] | | Outcome | ||
| 17 | Mean Ratio [Ache] | Calculated inhibitor-treated/DMSO ratio for Ache as mean for all unique peptides quantified | 6 | | Float | ratio | ||
| 18 | Standard Error [Ache] | Calculated standard error for mean | 6 | | Float | |||
| 19 | Outcome [Acot1] | One of Active, Inactive, or Not Tested | 7 | acyl-coenzyme A thioesterase 1 [Mus musculus] | | Outcome | ||
| 20 | Mean Ratio [Acot1] | Calculated inhibitor-treated/DMSO ratio for Acot1 as mean for all unique peptides quantified | 7 | | Float | ratio | ||
| 21 | Standard Error [Acot1] | Calculated standard error for mean | 7 | | Float | |||
| 22 | Outcome [Acot2] | One of Active, Inactive, or Not Tested | 8 | acyl-coenzyme A thioesterase 2, mitochondrial precursor [Mus musculus] | | Outcome | ||
| 23 | Mean Ratio [Acot2] | Calculated inhibitor-treated/DMSO ratio for Acot2 as mean for all unique peptides quantified | 8 | | Float | ratio | ||
| 24 | Standard Error [Acot2] | Calculated standard error for mean | 8 | | Float | |||
| 25 | Outcome [Acot7] | One of Active, Inactive, or Not Tested | 9 | cytosolic acyl coenzyme A thioester hydrolase isoform 1 [Mus musculus] | | Outcome | ||
| 26 | Mean Ratio [Acot7] | Calculated inhibitor-treated/DMSO ratio for Acot7 as mean for all unique peptides quantified | 9 | | Float | ratio | ||
| 27 | Standard Error [Acot7] | Calculated standard error for mean | 9 | | Float | |||
| 28 | Outcome [Acot9] | One of Active, Inactive, or Not Tested | 10 | acyl-coenzyme A thioesterase 9, mitochondrial [Mus musculus] | | Outcome | ||
| 29 | Mean Ratio [Acot9] | Calculated inhibitor-treated/DMSO ratio for Acot9 as mean for all unique peptides quantified | 10 | | Float | ratio | ||
| 30 | Standard Error [Acot9] | Calculated standard error for mean | 10 | | Float | |||
| 31 | Outcome [Apeh] | One of Active, Inactive, or Not Tested | 11 | Acylpeptide hydrolase [Mus musculus] | | Outcome | ||
| 32 | Mean Ratio [Apeh] | Calculated inhibitor-treated/DMSO ratio for Apeh as mean for all unique peptides quantified | 11 | | Float | ratio | ||
| 33 | Standard Error [Apeh] | Calculated standard error for mean | 11 | | Float | |||
| 34 | Outcome [Bat5] | One of Active, Inactive, or Not Tested | 12 | abhydrolase domain-containing protein 16A [Mus musculus] | | Outcome | ||
| 35 | Mean Ratio [Bat5] | Calculated inhibitor-treated/DMSO ratio for Bat5 as mean for all unique peptides quantified | 12 | | Float | ratio | ||
| 36 | Standard Error [Bat5] | Calculated standard error for mean | 12 | | Float | |||
| 37 | Outcome [Ctsa] | One of Active, Inactive, or Not Tested | 13 | lysosomal protective protein isoform b [Mus musculus] | | Outcome | ||
| 38 | Mean Ratio [Ctsa] | Calculated inhibitor-treated/DMSO ratio for Ctsa as mean for all unique peptides quantified | 13 | | Float | ratio | ||
| 39 | Standard Error [Ctsa] | Calculated standard error for mean | 13 | | Float | |||
| 40 | Outcome [Ddhd2] | One of Active, Inactive, or Not Tested | 14 | phospholipase DDHD2 [Mus musculus] | | Outcome | ||
| 41 | Mean Ratio [Ddhd2] | Calculated inhibitor-treated/DMSO ratio for Ddhd2 as mean for all unique peptides quantified | 14 | | Float | ratio | ||
| 42 | Standard Error [Ddhd2] | Calculated standard error for mean | 14 | | Float | |||
| 43 | Outcome [Dpp7] | One of Active, Inactive, or Not Tested | 15 | dipeptidyl peptidase 2 precursor [Mus musculus] | | Outcome | ||
| 44 | Mean Ratio [Dpp7] | Calculated inhibitor-treated/DMSO ratio for Dpp7 as mean for all unique peptides quantified | 15 | | Float | ratio | ||
| 45 | Standard Error [Dpp7] | Calculated standard error for mean | 15 | | Float | |||
| 46 | Outcome [Dpp8] | One of Active, Inactive, or Not Tested | 16 | dipeptidyl peptidase 8 [Mus musculus] | | Outcome | ||
| 47 | Mean Ratio [Dpp8] | Calculated inhibitor-treated/DMSO ratio for Dpp8 as mean for all unique peptides quantified | 16 | | Float | ratio | ||
| 48 | Standard Error [Dpp8] | Calculated standard error for mean | 16 | | Float | |||
| 49 | Outcome [Dpp9] | One of Active, Inactive, or Not Tested | 17 | dipeptidyl peptidase 9 [Mus musculus] | | Outcome | ||
| 50 | Mean Ratio [Dpp9] | Calculated inhibitor-treated/DMSO ratio for Dpp9 as mean for all unique peptides quantified | 17 | | Float | ratio | ||
| 51 | Standard Error [Dpp9] | Calculated standard error for mean | 17 | | Float | |||
| 52 | Outcome [Faah] | One of Active, Inactive, or Not Tested | 18 | fatty acid amide hydrolase [Mus musculus] | | Outcome | ||
| 53 | Mean Ratio [Faah] | Calculated inhibitor-treated/DMSO ratio for Faah as mean for all unique peptides quantified | 18 | | Float | ratio | ||
| 54 | Standard Error [Faah] | Calculated standard error for mean | 18 | | Float | |||
| 55 | Outcome [Fam108b1] | One of Active, Inactive, or Not Tested | 19 | abhydrolase domain-containing protein FAM108B1 precursor [Mus musculus] | | Outcome | ||
| 56 | Mean Ratio [Fam108b1] | Calculated inhibitor-treated/DMSO ratio for Fam108b1 as mean for all unique peptides quantified | 19 | | Float | ratio | ||
| 57 | Standard Error [Fam108b1] | Calculated standard error for mean | 19 | | Float | |||
| 58 | Outcome [Fasn] | One of Active, Inactive, or Not Tested | 20 | fatty acid synthase [Mus musculus] | | Outcome | ||
| 59 | Mean Ratio [Fasn] | Calculated inhibitor-treated/DMSO ratio for Fasn as mean for all unique peptides quantified | 20 | | Float | ratio | ||
| 60 | Standard Error [Fasn] | Calculated standard error for mean | 20 | | Float | |||
| 61 | Outcome [Htra2] | One of Active, Inactive, or Not Tested | 21 | serine protease HTRA2, mitochondrial [Mus musculus] | | Outcome | ||
| 62 | Mean Ratio [Htra2] | Calculated inhibitor-treated/DMSO ratio for Htra2 as mean for all unique peptides quantified | 21 | | Float | ratio | ||
| 63 | Standard Error [Htra2] | Calculated standard error for mean | 21 | | Float | |||
| 64 | Outcome [Iah1] | One of Active, Inactive, or Not Tested | 22 | isoamyl acetate-hydrolyzing esterase 1 homolog [Mus musculus] | | Outcome | ||
| 65 | Mean Ratio [Iah1] | Calculated inhibitor-treated/DMSO ratio for Iah1 as mean for all unique peptides quantified | 22 | | Float | ratio | ||
| 66 | Standard Error [Iah1] | Calculated standard error for mean | 22 | | Float | |||
| 67 | Outcome [Lypla1] | One of Active, Inactive, or Not Tested | 23 | acyl-protein thioesterase 1 [Mus musculus] | | Outcome | ||
| 68 | Mean Ratio [Lypla1] | Calculated inhibitor-treated/DMSO ratio for Lypla1 as mean for all unique peptides quantified | 23 | | Float | ratio | ||
| 69 | Standard Error [Lypla1] | Calculated standard error for mean | 23 | | Float | |||
| 70 | Outcome [Lypla2] | One of Active, Inactive, or Not Tested | 24 | acyl-protein thioesterase 2 [Mus musculus] | | Outcome | ||
| 71 | Mean Ratio [Lypla2] | Calculated inhibitor-treated/DMSO ratio for Lypla2 as mean for all unique peptides quantified | 24 | | Float | ratio | ||
| 72 | Standard Error [Lypla2] | Calculated standard error for mean | 24 | | Float | |||
| 73 | Outcome [Lyplal1] | One of Active, Inactive, or Not Tested | 25 | lysophospholipase-like protein 1 [Mus musculus] | | Outcome | ||
| 74 | Mean Ratio [Lyplal1] | Calculated inhibitor-treated/DMSO ratio for Lyplal1 as mean for all unique peptides quantified | 25 | | Float | ratio | ||
| 75 | Standard Error [Lyplal1] | Calculated standard error for mean | 25 | | Float | |||
| 76 | Outcome [Nceh1] | One of Active, Inactive, or Not Tested | 26 | neutral cholesterol ester hydrolase 1 [Mus musculus] | | Outcome | ||
| 77 | Mean Ratio [Nceh1] | Calculated inhibitor-treated/DMSO ratio for Nceh1 as mean for all unique peptides quantified | 26 | | Float | ratio | ||
| 78 | Standard Error [Nceh1] | Calculated standard error for mean | 26 | | Float | |||
| 79 | Outcome [Pafah1b2] | One of Active, Inactive, or Not Tested | 27 | platelet-activating factor acetylhydrolase IB subunit beta [Mus musculus] | | Outcome | ||
| 80 | Mean Ratio [Pafah1b2] | Calculated inhibitor-treated/DMSO ratio for Pafah1b2 as mean for all unique peptides quantified | 27 | | Float | ratio | ||
| 81 | Standard Error [Pafah1b2] | Calculated standard error for mean | 27 | | Float | |||
| 82 | Outcome [Pafah1b3] | One of Active, Inactive, or Not Tested | 28 | platelet-activating factor acetylhydrolase IB subunit gamma [Mus musculus] | | Outcome | ||
| 83 | Mean Ratio [Pafah1b3] | Calculated inhibitor-treated/DMSO ratio for Pafah1b3 as mean for all unique peptides quantified | 28 | | Float | ratio | ||
| 84 | Standard Error [Pafah1b3] | Calculated standard error for mean | 28 | | Float | |||
| 85 | Outcome [Pafah2] | One of Active, Inactive, or Not Tested | 29 | platelet-activating factor acetylhydrolase 2, cytoplasmic [Mus musculus] | | Outcome | ||
| 86 | Mean Ratio [Pafah2] | Calculated inhibitor-treated/DMSO ratio for Pafah2 as mean for all unique peptides quantified | 29 | | Float | ratio | ||
| 87 | Standard Error [Pafah2] | Calculated standard error for mean | 29 | | Float | |||
| 88 | Outcome [Parl] | One of Active, Inactive, or Not Tested | 30 | presenilins-associated rhomboid-like protein, mitochondrial precursor [Mus musculus] | | Outcome | ||
| 89 | Mean Ratio [Parl] | Calculated inhibitor-treated/DMSO ratio for Parl as mean for all unique peptides quantified | 30 | | Float | ratio | ||
| 90 | Standard Error [Parl] | Calculated standard error for mean | 30 | | Float | |||
| 91 | Outcome [Pgap1] | One of Active, Inactive, or Not Tested | 31 | GPI inositol-deacylase [Mus musculus] | | Outcome | ||
| 92 | Mean Ratio [Pgap1] | Calculated inhibitor-treated/DMSO ratio for Pgap1 as mean for all unique peptides quantified | 31 | | Float | ratio | ||
| 93 | Standard Error [Pgap1] | Calculated standard error for mean | 31 | | Float | |||
| 94 | Outcome [Pla2g15] | One of Active, Inactive, or Not Tested | 32 | group XV phospholipase A2 precursor [Mus musculus] | | Outcome | ||
| 95 | Mean Ratio [Pla2g15] | Calculated inhibitor-treated/DMSO ratio for Pla2g15 as mean for all unique peptides quantified | 32 | | Float | ratio | ||
| 96 | Standard Error [Pla2g15] | Calculated standard error for mean | 32 | | Float | |||
| 97 | Outcome [Pla2g6] | One of Active, Inactive, or Not Tested | 33 | 85 kDa calcium-independent phospholipase A2 isoform 2 [Mus musculus] | | Outcome | ||
| 98 | Mean Ratio [Pla2g6] | Calculated inhibitor-treated/DMSO ratio for Pla2g6 as mean for all unique peptides quantified | 33 | | Float | ratio | ||
| 99 | Standard Error [Pla2g6] | Calculated standard error for mean | 33 | | Float | |||
| 100 | Outcome [Pnpla8] | One of Active, Inactive, or Not Tested | 34 | calcium-independent phospholipase A2-gamma [Mus musculus] | | Outcome | ||
| 101 | Mean Ratio [Pnpla8] | Calculated inhibitor-treated/DMSO ratio for Pnpla8 as mean for all unique peptides quantified | 34 | | Float | ratio | ||
| 102 | Standard Error [Pnpla8] | Calculated standard error for mean | 34 | | Float | |||
| 103 | Outcome [Ppt2] | One of Active, Inactive, or Not Tested | 35 | lysosomal thioesterase PPT2 precursor [Mus musculus] | | Outcome | ||
| 104 | Mean Ratio [Ppt2] | Calculated inhibitor-treated/DMSO ratio for Ppt2 as mean for all unique peptides quantified | 35 | | Float | ratio | ||
| 105 | Standard Error [Ppt2] | Calculated standard error for mean | 35 | | Float | |||
| 106 | Outcome [Prcp] | One of Active, Inactive, or Not Tested | 36 | lysosomal Pro-X carboxypeptidase precursor [Mus musculus] | | Outcome | ||
| 107 | Mean Ratio [Prcp] | Calculated inhibitor-treated/DMSO ratio for Prcp as mean for all unique peptides quantified | 36 | | Float | ratio | ||
| 108 | Standard Error [Prcp] | Calculated standard error for mean | 36 | | Float | |||
| 109 | Outcome [Prep] | One of Active, Inactive, or Not Tested | 37 | prolyl endopeptidase [Mus musculus] | | Outcome | ||
| 110 | Mean Ratio [Prep] | Calculated inhibitor-treated/DMSO ratio for Prep as mean for all unique peptides quantified | 37 | | Float | ratio | ||
| 111 | Standard Error [Prep] | Calculated standard error for mean | 37 | | Float | |||
| 112 | Outcome [Prepl] | One of Active, Inactive, or Not Tested | 38 | prolyl endopeptidase-like isoform a [Mus musculus] | | Outcome | ||
| 113 | Mean Ratio [Prepl] | Calculated inhibitor-treated/DMSO ratio for Prepl as mean for all unique peptides quantified | 38 | | Float | ratio | ||
| 114 | Standard Error [Prepl] | Calculated standard error for mean | 38 | | Float | |||
| 115 | Outcome [Rbbp9] | One of Active, Inactive, or Not Tested | 39 | putative hydrolase RBBP9 [Mus musculus] | | Outcome | ||
| 116 | Mean Ratio [Rbbp9] | Calculated inhibitor-treated/DMSO ratio for Rbbp9 as mean for all unique peptides quantified | 39 | | Float | ratio | ||
| 117 | Standard Error [Rbbp9] | Calculated standard error for mean | 39 | | Float | |||
| 118 | Outcome [Serhl] | One of Active, Inactive, or Not Tested | 40 | serine hydrolase-like protein [Mus musculus] | | Outcome | ||
| 119 | Mean Ratio [Serhl] | Calculated inhibitor-treated/DMSO ratio for Serh1 as mean for all unique peptides quantified | 40 | | Float | ratio | ||
| 120 | Standard Error [Serhl] | Calculated standard error for mean | 40 | | Float |
Additional Information
Grant Number: 1 R01 CA132630-01
Classification
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